{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T04:58:21Z","timestamp":1770267501630,"version":"3.49.0"},"reference-count":51,"publisher":"Portland Press Ltd.","issue":"1","license":[{"start":{"date-parts":[[2020,12,8]],"date-time":"2020-12-08T00:00:00Z","timestamp":1607385600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/2.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2021,1,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Chronic allograft dysfunction is a major cause of late graft failure after kidney transplantation. One of the histological changes is interstitial fibrosis, which is associated with epithelial\u2013mesenchymal transition. Bortezomib has been reported to prevent the progression of fibrosis in organs. We used rat renal transplantation model and human kidney 2 cell line treated with tumor necrosis factor-\u03b1 (TNF-\u03b1) to examine their response to bortezomib. To explore the mechanism behind it, we assessed the previously studied TNF-\u03b1\/protein kinase B (Akt)\/Smad ubiquitin regulatory factor 2 (Smurf2) signaling and performed RNA sequencing. Our results suggested that bortezomib could attenuate the TNF-\u03b1-induced epithelial\u2013mesenchymal transition and renal allograft interstitial fibrosis in vitro and in vivo. In addition to blocking Akt\/mammalian target of rapamycin (mTOR)\/p70S6 kinase\/Smurf2 signaling, bortezomib\u2019s effect on the epithelial\u2013mesenchymal transition was associated with inhibition of nuclear factor kappa B (NF-\u03baB) pathway by stabilizing inhibitor of NF-\u03baB. The study highlighted the therapeutic potential of bortezomib on renal allograft interstitial fibrosis. Such an effect may result from inhibition of NF-\u03baB\/TNF-\u03b1\/Akt\/mTOR\/p70S6 kinase\/Smurf2 signaling via stabilizing protein of inhibitor of NF-\u03baB.<\/jats:p>","DOI":"10.1042\/cs20201038","type":"journal-article","created":{"date-parts":[[2020,12,9]],"date-time":"2020-12-09T22:12:34Z","timestamp":1607551954000},"page":"53-69","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Bortezomib limits renal allograft interstitial fibrosis by inhibiting NF-\u03baB\/TNF-\u03b1\/Akt\/mTOR\/P70S6K\/Smurf2 pathway via I\u03baB\u03b1 protein stabilization"],"prefix":"10.1042","volume":"135","author":[{"given":"Chuanjian","family":"Suo","sequence":"first","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zeping","family":"Gui","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zijie","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiajun","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hao","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuang","family":"Fei","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min","family":"Gu","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruoyun","family":"Tan","sequence":"additional","affiliation":[{"name":"Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2021,1,5]]},"reference":[{"key":"2021113011180880100_B1","doi-asserted-by":"publisher","first-page":"e46","DOI":"10.1097\/TP.0000000000003012","article-title":"The Causes of Kidney Allograft Failure: More Than Alloimmunity. A Viewpoint Article","volume":"104","author":"Van Loon","year":"2020","journal-title":"Transplantation"},{"key":"2021113011180880100_B2","doi-asserted-by":"publisher","first-page":"409","DOI":"10.21037\/atm.2018.09.68","article-title":"Solid organ transplantation in the 21(st) century","volume":"6","author":"Black","year":"2018","journal-title":"Ann. Transl. Med."},{"key":"2021113011180880100_B3","doi-asserted-by":"publisher","first-page":"450","DOI":"10.1111\/j.1600-6143.2010.03283.x","article-title":"Long-term renal allograft survival in the United States: a critical reappraisal","volume":"11","author":"Lamb","year":"2011","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B4","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1111\/ajt.14124","article-title":"OPTN\/SRTR 2015 Annual Data Report: Kidney","volume":"17","author":"Hart","year":"2017","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B5","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1111\/ajt.13423","article-title":"Changing Kidney Allograft Histology Early Posttransplant: Prognostic Implications of 1-Year Protocol Biopsies","volume":"16","author":"Cosio","year":"2016","journal-title":"Am. J. Transplant.: Off. J. Am Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B6","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1007\/978-981-13-8871-2_6","article-title":"Pathogenesis of Chronic Allograft Dysfunction Progress to Renal Fibrosis","volume":"1165","author":"Yang","year":"2019","journal-title":"Adv. Exp. Med. Biol."},{"key":"2021113011180880100_B7","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1146\/annurev-physiol-022516-034227","article-title":"Mechanisms of Renal Fibrosis","volume":"80","author":"Humphreys","year":"2018","journal-title":"Annu. Rev. Physiol."},{"key":"2021113011180880100_B8","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1016\/j.cell.2016.06.028","article-title":"EMT: 2016","volume":"166","author":"Nieto","year":"2016","journal-title":"Cell"},{"key":"2021113011180880100_B9","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/j.mehy.2013.03.020","article-title":"Epithelial-mesenchymal transition of renal tubules: divergent processes of repairing in acute or chronic injury?","volume":"81","author":"Jiang","year":"2013","journal-title":"Med. Hypotheses."},{"key":"2021113011180880100_B10","doi-asserted-by":"publisher","first-page":"3482","DOI":"10.1016\/j.transproceed.2010.06.014","article-title":"Vimentin expression and myofibroblast infiltration are early markers of renal dysfunction in kidney transplantation: an early stage of chronic allograft dysfunction?","volume":"42","author":"de Matos","year":"2010","journal-title":"Transplant. Proc."},{"key":"2021113011180880100_B11","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1038\/ki.2014.285","article-title":"TNF receptors: signaling pathways and contribution to renal dysfunction","volume":"87","author":"Al-Lamki","year":"2015","journal-title":"Kidney Int."},{"key":"2021113011180880100_B12","doi-asserted-by":"publisher","first-page":"e12502","DOI":"10.1111\/xen.12502","article-title":"Potential pathological role of pro-inflammatory cytokines (IL-6, TNF-alpha, and IL-17) in xenotransplantation","volume":"26","author":"Zhao","year":"2019","journal-title":"Xenotransplantation"},{"key":"2021113011180880100_B13","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1007\/s13402-019-00454-y","article-title":"The role of TNF-alpha in chordoma progression and inflammatory pathways","volume":"42","author":"Gulluoglu","year":"2019","journal-title":"Cell. Oncol. (Dordrecht)"},{"key":"2021113011180880100_B14","doi-asserted-by":"publisher","first-page":"Cd010816","DOI":"10.1002\/14651858.CD010816.pub2","article-title":"Bortezomib for the treatment of multiple myeloma","volume":"4","author":"Scott","year":"2016","journal-title":"Cochrane Database Systematic Rev."},{"key":"2021113011180880100_B15","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1007\/s00535-012-0675-z","article-title":"Bortezomib induces tumor-specific cell death and growth inhibition in hepatocellular carcinoma and improves liver fibrosis","volume":"48","author":"Saeki","year":"2013","journal-title":"J. Gastroenterol."},{"key":"2021113011180880100_B16","doi-asserted-by":"publisher","first-page":"13086","DOI":"10.1038\/s41598-017-13486-x","article-title":"The proteasome inhibitor bortezomib attenuates renal fibrosis in mice via the suppression of TGF-beta1","volume":"7","author":"Zeniya","year":"2017","journal-title":"Sci. Rep."},{"key":"2021113011180880100_B17","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1161\/01.HYP.0000142772.71367.65","article-title":"Downregulation of matrix metalloproteinases and collagens and suppression of cardiac fibrosis by inhibition of the proteasome","volume":"44","author":"Meiners","year":"2004","journal-title":"Hypertension (Dallas, Tex.: 1979)"},{"key":"2021113011180880100_B18","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1182\/blood-2006-10-054502","article-title":"Proteasome inhibitor bortezomib impairs both myelofibrosis and osteosclerosis induced by high thrombopoietin levels in mice","volume":"110","author":"Wagner-Ballon","year":"2007","journal-title":"Blood"},{"key":"2021113011180880100_B19","doi-asserted-by":"publisher","first-page":"3764","DOI":"10.1093\/ndt\/gfq230","article-title":"Bortezomib and sirolimus inhibit the chronic active antibody-mediated rejection in experimental renal transplantation in the rat","volume":"25","author":"Vogelbacher","year":"2010","journal-title":"Nephrol. Dialysis Transplant.: Off. Publ. Eur. Dialysis Transplant Assoc. - Eur. Renal Assoc."},{"key":"2021113011180880100_B20","doi-asserted-by":"publisher","first-page":"2916","DOI":"10.1097\/01.ASN.0000034909.10994.8A","article-title":"Pentoxifylline attenuated the renal disease progression in rats with remnant kidney","volume":"13","author":"Lin","year":"2002","journal-title":"J. Am. Soc. Nephrol.: JASN"},{"key":"2021113011180880100_B21","doi-asserted-by":"publisher","first-page":"2702","DOI":"10.1681\/ASN.2005040435","article-title":"Pentoxifylline attenuates tubulointerstitial fibrosis by blocking Smad3\/4-activated transcription and profibrogenic effects of connective tissue growth factor","volume":"16","author":"Lin","year":"2005","journal-title":"J. Am. Soc. Nephrol.: JASN"},{"key":"2021113011180880100_B22","doi-asserted-by":"crossref","first-page":"1542","DOI":"10.1681\/ASN.V1071542","article-title":"Combining an antiproteinuric approach with mycophenolate mofetil fully suppresses progressive nephropathy of experimental animals","volume":"10","author":"Remuzzi","year":"1999","journal-title":"J. Am. Soc. Nephrol.: JASN"},{"key":"2021113011180880100_B23","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1007\/978-981-13-8871-2_18","article-title":"Inflammatory Mediators and Renal Fibrosis","volume":"1165","author":"Meng","year":"2019","journal-title":"Adv. Exp. Med. Biol."},{"key":"2021113011180880100_B24","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1369\/0022155419852932","article-title":"Renal Inflammation and Fibrosis: A Double-edged Sword","volume":"67","author":"Black","year":"2019","journal-title":"J. Histochem. Cytochem.: Off. J. Histochem. Soc."},{"key":"2021113011180880100_B25","doi-asserted-by":"publisher","first-page":"461","DOI":"10.1016\/j.juro.2008.04.001","article-title":"Cytokines in epithelial-mesenchymal transition: a new insight into obstructive nephropathy","volume":"180","author":"Bani-Hani","year":"2008","journal-title":"J. Urol."},{"key":"2021113011180880100_B26","doi-asserted-by":"publisher","first-page":"646","DOI":"10.1016\/j.kint.2017.02.012","article-title":"Antagonism of profibrotic microRNA-21 improves outcome of murine chronic renal allograft dysfunction","volume":"92","author":"Schauerte","year":"2017","journal-title":"Kidney Int."},{"key":"2021113011180880100_B27","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1111\/ajt.14431","article-title":"Renal Allograft Histology at 10 Years After Transplantation in the Tacrolimus Era: Evidence of Pervasive Chronic Injury","volume":"18","author":"Stegall","year":"2018","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B28","doi-asserted-by":"publisher","first-page":"85","DOI":"10.2353\/ajpath.2010.090517","article-title":"Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis","volume":"176","author":"Humphreys","year":"2010","journal-title":"Am. J. Pathol."},{"key":"2021113011180880100_B29","doi-asserted-by":"publisher","first-page":"1767","DOI":"10.2353\/ajpath.2010.090345","article-title":"Autophagy is a component of epithelial cell fate in obstructive uropathy","volume":"176","author":"Li","year":"2010","journal-title":"Am. J. Pathol."},{"key":"2021113011180880100_B30","doi-asserted-by":"publisher","first-page":"1047","DOI":"10.1038\/nm.3218","article-title":"Origin and function of myofibroblasts in kidney fibrosis","volume":"19","author":"LeBleu","year":"2013","journal-title":"Nat. Med."},{"key":"2021113011180880100_B31","doi-asserted-by":"publisher","first-page":"468","DOI":"10.1172\/JCI44595","article-title":"Epithelial-mesenchymal transition (EMT) in kidney fibrosis: fact or fantasy?","volume":"121","author":"Kriz","year":"2011","journal-title":"J. Clin. Invest."},{"key":"2021113011180880100_B32","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1111\/j.1365-2613.2011.00775.x","article-title":"Epithelial-mesenchymal transition in renal fibrosis - evidence for and against","volume":"92","author":"Fragiadaki","year":"2011","journal-title":"Int. J. Exp. Pathol."},{"key":"2021113011180880100_B33","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1016\/j.gene.2017.11.059","article-title":"Role of tumor necrosis factor-alpha in epithelial-to-mesenchymal transition in transplanted kidney cells in recipients with chronic allograft dysfunction","volume":"642","author":"Zhao","year":"2018","journal-title":"Gene"},{"key":"2021113011180880100_B34","doi-asserted-by":"publisher","first-page":"1674","DOI":"10.1002\/hep.30706","article-title":"Proteasomal Degradation of Enhancer of Zeste Homologue 2 in Cholangiocytes Promotes Biliary Fibrosis","volume":"70","author":"Jalan-Sakrikar","year":"2019","journal-title":"Hepatology"},{"key":"2021113011180880100_B35","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1016\/j.yjmcc.2014.12.007","article-title":"Activation of the cardiac proteasome promotes angiotension II-induced hypertrophy by down-regulation of ATRAP","volume":"79","author":"Li","year":"2015","journal-title":"J. Mol. Cell Cardiol."},{"key":"2021113011180880100_B36","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.trim.2013.12.003","article-title":"Protective role of bortezomib in steatotic liver ischemia\/reperfusion injury through abrogation of MMP activation and YKL-40 expression","volume":"30","author":"Tiriveedhi","year":"2014","journal-title":"Transpl. Immunol."},{"key":"2021113011180880100_B37","doi-asserted-by":"publisher","first-page":"751","DOI":"10.1016\/j.ijrobp.2016.01.021","article-title":"Proteomic Profiling of Radiation-Induced Skin Fibrosis in Rats: Targeting the Ubiquitin-Proteasome System","volume":"95","author":"Wang","year":"2016","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"2021113011180880100_B38","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1513\/pats.200906-055JS","article-title":"Proteasomal regulation of pulmonary fibrosis","volume":"7","author":"Weiss","year":"2010","journal-title":"Proc. Am. Thoracic Soc."},{"key":"2021113011180880100_B39","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.cell.2016.12.012","article-title":"30 Years of NF-kappaB: A Blossoming of Relevance to Human Pathobiology","volume":"168","author":"Zhang","year":"2017","journal-title":"Cell"},{"key":"2021113011180880100_B40","first-page":"309","article-title":"NF-kappaB, inflammation, immunity and cancer: coming of age. Nature reviews","volume":"18","author":"Taniguchi","year":"2018","journal-title":"Immunology"},{"key":"2021113011180880100_B41","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1016\/j.it.2016.06.002","article-title":"More to Life than NF-kappaB in TNFR1 Signaling","volume":"37","author":"Ting","year":"2016","journal-title":"Trends Immunol."},{"key":"2021113011180880100_B42","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1038\/ki.2014.287","article-title":"Origin of myofibroblasts and cellular events triggering fibrosis","volume":"87","author":"Mack","year":"2015","journal-title":"Kidney Int."},{"key":"2021113011180880100_B43","doi-asserted-by":"publisher","first-page":"8936856","DOI":"10.1155\/2019\/8936856","article-title":"Fenofibrate Improved Interstitial Fibrosis of Renal Allograft through Inhibited Epithelial-Mesenchymal Transition Induced by Oxidative Stress","volume":"2019","author":"Wang","year":"2019","journal-title":"Oxidative Med. Cell. Longevity"},{"key":"2021113011180880100_B44","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1159\/000487501","article-title":"The Association Between Oxidative Stress Alleviation via Sulforaphane-Induced Nrf2-HO-1\/NQO-1 Signaling Pathway Activation and Chronic Renal Allograft Dysfunction Improvement","volume":"43","author":"Lv","year":"2018","journal-title":"Kidney Blood Pressure Res."},{"key":"2021113011180880100_B45","doi-asserted-by":"publisher","first-page":"2223","DOI":"10.1111\/j.1600-6143.2009.02762.x","article-title":"Wnt pathway regulation in chronic renal allograft damage","volume":"9","author":"von Toerne","year":"2009","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B46","doi-asserted-by":"publisher","first-page":"3696","DOI":"10.3892\/mmr.2019.10045","article-title":"SDF-1\/CXCR4 induces epithelial-mesenchymal transition through activation of the Wnt\/\u03b2-catenin signaling pathway in rat chronic allograft nephropathy","volume":"19","author":"Tang","year":"2019","journal-title":"Mol. Med. Rep."},{"key":"2021113011180880100_B47","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1111\/j.1600-6143.2011.03776.x","article-title":"Modulation of Wnt and Hedgehog signaling pathways is linked to retinoic acid-induced amelioration of chronic allograft dysfunction","volume":"12","author":"von Toerne","year":"2012","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"},{"key":"2021113011180880100_B48","doi-asserted-by":"publisher","DOI":"10.3390\/nu11040722","article-title":"Lipid accumulation and chronic kidney disease","volume":"11","author":"Gai","year":"2019","journal-title":"Nutrients"},{"key":"2021113011180880100_B49","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1097\/00007890-199702150-00001","article-title":"Hyperlipidemia in solid organ transplantation","volume":"63","author":"Kobashigawa","year":"1997","journal-title":"Transplantation"},{"key":"2021113011180880100_B50","doi-asserted-by":"publisher","first-page":"1467","DOI":"10.1681\/ASN.2011100967","article-title":"Cellular senescence limits regenerative capacity and allograft survival","volume":"23","author":"Braun","year":"2012","journal-title":"J. Am. Soc. Nephrol.: JASN"},{"key":"2021113011180880100_B51","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1111\/j.1600-6143.2008.02500.x","article-title":"Effects of donor age and cell senescence on kidney allograft survival","volume":"9","author":"Melk","year":"2009","journal-title":"Am. J. Transplant.: Off. J. Am. Soc. Transplant. Am. Soc. Transplant Surgeons"}],"container-title":["Clinical Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/clinsci\/article-pdf\/135\/1\/53\/900976\/cs-2020-1038.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/clinsci\/article-pdf\/135\/1\/53\/900976\/cs-2020-1038.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T12:16:03Z","timestamp":1638274563000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/clinsci\/article\/135\/1\/53\/227193\/Bortezomib-limits-renal-allograft-interstitial"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1]]},"references-count":51,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,1,15]]}},"URL":"https:\/\/doi.org\/10.1042\/cs20201038","relation":{},"ISSN":["0143-5221","1470-8736"],"issn-type":[{"value":"0143-5221","type":"print"},{"value":"1470-8736","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2021,1]]},"published":{"date-parts":[[2021,1]]}}}